title: Non-viral transfection methods optimized for gene delivery to a lung cancer cell line.
abstract: BACKGROUND
  Mehr-80 is a newly established adherent human large cell lung cancer cell line that has not been transfected until now. This study aims to define the optimal transfection conditions and effects of some critical elements for enhancing gene delivery to this cell line by utilizing different non-viral transfection Procedures.
  
  
  METHODS
  In the current study, calcium phosphate (CaP), DEAE-dextran, superfect, electroporation and lipofection transfection methods were used to optimize delivery of a plasmid construct that expressed Green Fluorescent Protein (GFP). Transgene expression was detected by fluorescent microscopy and flowcytometry. Toxicities of the methods were estimated by trypan blue staining. In order to evaluate the density of the transfected gene, we used a plasmid construct that expressed the Stromal cell-Derived Factor-1 (SDF-1) gene and measured its expression by real-time PCR.
  
  
  RESULTS
  Mean levels of GFP-expressing cells 48 hr after transfection were 8.4% (CaP), 8.2% (DEAE-dextran), 4.9% (superfect), 34.1% (electroporation), and 40.1% (lipofection). Lipofection had the highest intense SDF-1 expression of the analyzed methods.
  
  
  CONCLUSIONS
  This study has shown that the lipofection and electroporation methods were more efficient at gene delivery to Mehr-80 cells. The quantity of DNA per transfection, reagent concentration, and incubation time were identified as essential factors for successful transfection in all of the studied methods.
authors.affiliation: Shiraz Institute for Cancer Research, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: Loghman
authors.initials: L
authors.lastname: Salimzadeh
authors.forename: Mansooreh
authors.initials: M
authors.lastname: Jaberipour
authors.forename: Ahmad
authors.initials: A
authors.lastname: Hosseini
authors.forename: Abbas
authors.initials: A
authors.lastname: Ghaderi
keywords: Cell line; Green flourescent proteins; Lung cancer; Transfection